Escherichia coli ribosomes as a model for testing new photoactivated tRNA analogs containing 6-thioguanosine residues

I. N. Lavrik, P. V. Sergiev, A. A. Bogdanov, R. A. Zimmermann, O. A. Dontsova

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

A comparative study of photoreactive tRNA derivatives containing a 6-thioguanosine or diazirine derivative of 5-methyleneaminouridine was performed using Escherichia coli ribosomes. The conditions of producing functional tRNAs containing photoreactive residues have been selected using T7 RNA polymerase transcription. Photochemical modification was conducted upon binding the corresponding tRNA Phe transcripts carrying the photoreactive residues of nucleotide analogs to the P site of the ribosome. The results of modification were analyzed using oligonucleotide-directed hydrolysis of rRNA by RNase H, as well as reverse transcription. It is shown that 6-thioguanosine is a promising agent for studying RNA-protein complexes by photochemical crosslinking methods.

Original languageEnglish
Pages (from-to)799-805
Number of pages7
JournalMolecular Biology
Volume38
Issue number5
DOIs
Publication statusPublished - Sep 2004
Externally publishedYes

Keywords

  • 6-thioguanosine
  • Escherichia coli
  • photoaffinity modification
  • ribosome
  • tRNA

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